Increasing PCR sensitivity by removal of polymerase inhibitors in environmental samples by using dielectrophoresis

被引:20
作者
del Carmen Jaramillo, Maria [1 ]
Martinez-Duarte, Rodrigo [2 ]
Huettener, Mario [1 ]
Renaud, Philippe [2 ]
Torrents, Eduard [1 ]
Juarez, Antonio [1 ,3 ]
机构
[1] Inst Bioengn Catalonia IBEC, Barcelona 08028, Spain
[2] Ecole Polytech Fed Lausanne, Microsyst Lab LMIS4, CH-1015 Lausanne, Switzerland
[3] Univ Barcelona, Dept Microbiol, Fac Biol, E-08028 Barcelona, Spain
关键词
Yeast; Dielectrophoresis; PCR; Separation; Carbon-electrode; CONTINUOUS SEPARATION; CELLS; MICROORGANISMS; PARTICLES; IMPEDANCE; MIXTURES;
D O I
10.1016/j.bios.2012.12.049
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Dielectrophoresis (DEP) is a powerful tool to manipulate cells and molecules in microfluidic chips. However, few practical applications using DEP exist. An immediate practical application of a carbon-electrode DEP system, in removing PCR inhibitors from a sample, is reported in this work. We use a high throughput carbon-electrode DEP system to trap yeast cells from a natural sample (fermented grape must) and then in situ remove contaminants that interfere with PCR analysis. Retrieval of this enriched and purified yeast population from the DEP system then allows for a significant increase of sensitivity during PCR analysis. Furthermore, the fact that DEP can discriminate between viable and non-viable cells minimizes the number of false positives commonly obtained when using PCR alone. Experimental results provide clear evidence that the carbon-electrode DEP-based sample preparation step can readily and effectively clean environmental samples from natural contaminants and improve PCR sensitivity. (c) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:297 / 303
页数:7
相关论文
共 51 条
[1]   Forensic implications of PCR inhibition-A review [J].
Alaeddini, Reza .
FORENSIC SCIENCE INTERNATIONAL-GENETICS, 2012, 6 (03) :297-305
[2]   Interdigitated comb-like electrodes for continuous separation of malignant cells from blood using dielectrophoresis [J].
Alazzam, Anas ;
Stiharu, Ion ;
Bhat, Rama ;
Meguerditchian, Ari-Nareg .
ELECTROPHORESIS, 2011, 32 (11) :1327-1336
[3]  
[Anonymous], ISRN ONCOL
[4]  
[Anonymous], 1978, CAMBRIDGE MONOGRAPHS
[5]   Discrimination between Bacillus species by impedance analysis of individual dielectrophoretically positioned spores [J].
Beck, Joseph D. ;
Shang, Lu ;
Li, Bo ;
Marcus, Matthew S. ;
Hamers, Robert J. .
ANALYTICAL CHEMISTRY, 2008, 80 (10) :3757-3761
[6]   PCR-based detection in a micro-fabricated platform [J].
Bhattacharya, Shantanu ;
Salamat, Shuaib ;
Morisette, Dallas ;
Banada, Padmapriya ;
Akin, Demir ;
Liu, Yi-Shao ;
Bhunia, Arun K. ;
Ladisch, Michael ;
Bashir, Rashid .
LAB ON A CHIP, 2008, 8 (07) :1130-1136
[7]  
Borgatti M, 2005, INT J MOL MED, V15, P913
[8]   Dielectrophoresis as a tool to characterize and differentiate isogenic mutants of Escherichia coli [J].
Castellarnau, M. ;
Errachid, A. ;
Madrid, C. ;
Juarez, A. ;
Samitier, J. .
BIOPHYSICAL JOURNAL, 2006, 91 (10) :3937-3945
[9]   Lab-on-a-chip device for continuous particle and cell separation based on electrical properties via alternating current dielectrophoresis [J].
Cetin, Barbaros ;
Li, Dongqing .
ELECTROPHORESIS, 2010, 31 (18) :3035-3043
[10]   Commercialization of microfluidic point-of-care diagnostic devices [J].
Chin, Curtis D. ;
Linder, Vincent ;
Sia, Samuel K. .
LAB ON A CHIP, 2012, 12 (12) :2118-2134